| Crosbi ID | 186371 |
| Bib IRB ID | 593635 |
| Year | 2013 |
| Status | Objavljeno |
| Type | prilog u časopisu |
| Subtype | izvorni znanstveni rad |
| Authors | Leščić Ašler, Ivana ; Kovačić, Filip ; Marchetti-Deschmann, Martina ; Allmaier, Günter ; Štefanić, Zoran ; Kojić-Prodić, Biserka |
| Journal | Journal of enzyme inhibition and medicinal chemistry |
| Volume | 28 |
| Issue | 5 |
| Pages | str. 1094-1104 |
| DOI | 10.3109/14756366.2012.716834 |
| ISSN | 1475-6366 |
| EISSN | 1475-6374 |
en original Inhibition of extracellular lipase from Streptomyces rimosus with 3, 4-dichloroisocoumarin
Kinetic characterization of lipase inhibition was performed by activity measurement and mass spectrometry (MS), for the first time with serine protease inhibitor 3, 4-dichloroisocoumarin (DCI). Inhibition of Streptomyces rimosus extracellular lipase (SrLip), a member of the SGNH superfamily, by means of DCI follows the mechanism of two-step irreversible inhibition. The dissociation constant of the noncovalent EI complex and first-order rate constant for inactivation were determined by incubation (Ki* = 26.6 ± 2.8 μM, k2 = 12.2 ± 0.6 min–1) or progress curve (Ki* = 6.5 ± 1.5 μM, k2 = 0.11 ± 0.01 min–1) method. Half-times of reactivation for lipase inhibited with 10-fold molar excess of DCI were determined by activity measurement (t1/2 = 11.3 ± 0.2 h), matrix-assisted laser desorption/ionization (MALDI, t1/2 = 13.5 ± 0.4 h), and electro-spray ionization (ESI, t1/2 = 12.2 ± 0.5 h) MS. The active SrLip concentration was determined by incubating the enzyme with near equimolar concentrations of DCI, followed by activity and MS measurement.
en original Enzyme inhibition kinetics; active-site titration; mass spectrometry; SGNH-hydrolase
Leščić Ašler, Ivana ; Kovačić, Filip ; Marchetti-Deschmann, Martina ; Allmaier, Günter ; Štefanić, Zoran ; Kojić-Prodić, Biserka Inhibition of extracellular lipase from Streptomyces rimosus with 3, 4-dichloroisocoumarin // Journal of enzyme inhibition and medicinal chemistry, 28 (2013), 5; 1094-1104. doi: 10.3109/14756366.2012.716834
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